Comprehensive immune profiling unravels evolution of spatial distribution and immune repertoire in tumor microenvironment from primary to metastatic triple-negative breast cancer.
Authors: Weihua Guo, Colt A. Egelston, Katherine E. Hutchinson, Jennifer M. Giltnane, Veronica Ibarra-Lopez, Bonnie Liu, Susan Elaine Yost, Daniel Schmolze, Peter P. Lee, Yuan Yuan
Journal: Journal of Clinical Oncology (2022)

1079 Background: Immune checkpoint inhibitors (ICI) have improved PFS and OS in metastatic triple-negative breast cancer (mTNBC), but benefit is limited to PD-L1 positive tumors. Metastatic tumors are notorious for deficient immune cell infiltration and immunosuppressive features that may limit responses to ICI in mTNBC. However, the underlying mechanisms for the weak immunogenicity of the metastatic tumor immune microenvironment (TIME) and related poor ICI responses are still not well understood. The current study was designed to investigate the evolution of the TIME between paired primary and metastatic TNBCs. Methods: Spatial distribution of 37 key immune regulators using the NanoString digital spatial profiling (DSP) platform was analyzed. 452 regions of interests (ROIs) from 33 primary tumors (PT) and 29 metastatic tumors (MT) including 28 paired specimens, were selected based on CD45+ immune hotspots, and the protein expression levels of the key immune regulators were quantified within pan-cytokeratin (panCK) and CD45 masked regions, respectively. In parallel, we examined the clonality of tumor infiltrating B cell receptors by reconstructing the immune repertoire from bulk RNA-seq data. Results: Using the DSP platform, we confirmed reduced immune infiltration (e.g., CD3 and CD20) in both panCK and CD45 masked regions of MT, while CD8A and CD11c (p adj = 2.8×10 -7 and 2.1×10 -6 ) expression was only observed in panCK masked regions of MT compared with PT. A significant shift in myeloid composition between PT and MT as evidenced by increased CD68 signal (p adj = 5.8×10 -4 ) in CD45 masked regions of MT was identified. Within MT, PD-L1 signal was substantially higher (p adj = 0.030) in CD45 masked regions only, while PD1 counts were lower (p adj = 0.035) in panCK masked regions. This suggests the limited responses to ICI for MT may stem from relatively low expression of activated and targetable T cell subsets in MT islands. In support of the lower CD20 counts in MT, immune repertoire analysis revealed B cell receptor (BCR) repertoire diversity (represented by Gini index) was substantially lower in MT than PT (p = 0.041) suggesting that the ability of B cells to recognize a wide variety of tumor antigens in MT is greatly reduced in contrast to PT. Conclusions: Through comprehensive analysis of the TIME spatial organization within paired PT and MT, a significant reduction in dendritic cell/macrophage ratios (CD11c/CD68), reduced tumor localized T cell activation (CD8, PD1, PD-L1), and reduced B cell diversity (BCR clonality) are key features of the reduced immunogenicity of the metastatic TIME in TNBC. Further work to understand key mechanistic features driving the evolution of these differences in TIME between primary and metastatic tumors are ongoing.